To compare the activity of RGD-TRAIL in different expression systems, RGD-TRAIL in both Escherichia coli (E.coli) and Pichia pastoris was constructed and expressed. In vitro activity of RGD-TRAIL from Pichia pastoris expression system was also analyzed. Genetic engineering techniques were used to construct recombinant plasmid pET30-rgd-trail and pHBM-rgd-trail. The recombinant protein RGD-TRAIL was purified with Ni ion affinity chromatography after induction. MTT assay, ELISA, scratch wound healing, transwell migration assay and Hoechst 33342 staining were performed to detect the effects of RGD-TRAIL on proliferation, binding activity, migration and apoptosis. The expression of apoptosis-associated proteins was detected by Western blotting. Recombinant protein RGD-TRAIL was successfully expressed in a form of inclusion body in E.coli, while expressed secretorily in Pichia pastoris. It possessed more potent cytotoxicity than RGD-TRAIL in E.coli by MTT assay. The RGD-TRAIL expressed by Pichia pastoris showed powerful binding affinity with cancer cells expressing α(v), DR4, DR5 and highly potent cytotoxicity through inducing apoptosis of cancer cells. Nuclear fragmentation was examined by Hoechst 33342 staining. Cleaved PARP and caspase-3 were also detected after incubation with RGD-TRAIL. Additionally, RGD-TRAIL inhibited migration significantly in A549 and HT1080 cells. The results demonstrate that Pichia pastoris expression system is more suitable for the recombinant protein RGD-TRAIL. Its binding affinity and antitumor activity might make RGD-TRAIL a promising candidate for cancer therapy.

OBJECTIVE To explore the relationship between the change in Toll-like receptor expression and prognosis of severe viral hapatitsis.METHODS The peripheral blood mononuclear cells were separated and the expression of TLR2 and TLR4 mRNA was observed by reverse transcription-polymerase chain reaction(RT-PCR).The level of serum endotoxin was measured by chromogenic substrate Limulus amebocyte lysate assay.The data were analyzed by SPSS 11.5 software.RESULTS The expression of TLR2 and TLR4 mRNA and the level of serum endotoxin of death group before therapy were lightly higher than that of survival group,but after therapy that of death group is higher than survival group(P

Objective To study the clinical effect of complex treatment combined with external application of Rhubarb and Mirabilite on abdomen of patients with abdominal distension after abdominal surgery .Methods 90 patients with abdominal distension after gynecological abdominal surgery from June 2014 to September 2015 were randomly divided into conventional group(45 patients) and comprehensive group(45 patients).Patients in the conventional group were given routine gynecological postoperative care, and patients in the comprehensive group were given complex treatment combined with external application of Rhubarb and Mirabilite on abdomen.The abdominal distension duration, anal exhaust time, time of resuming peristaltic sound, first defecation time, treatment effect, depression and anxiety emotions, changes of life quality of the two groups before and after intervention were compared.Results The comprehensive group had higher treatment effect than the comprehensive group ( P<0.05 ) .The abdominal distension duration, anal exhaust time, time of resuming peristaltic sound, first defecation time of the comprehensive group were shorter than that of the conventional group(P<0.05).The comprehensive group had better depression, anxiety emotions and life quality compared to the conventional group (P<0.05).Conclusion Complex treatment combined with external application of Rhubarb and Mirabilite on abdomen has a very significant clinical effect on patients with abdominal distension after abdominal surgery, it can improve patients’negative emotions and the symptom of abdomen distension, promote life quality.

OBJECTIVE To investigate the clinical and etiology characteristics of nosocomial pneumonia. METHODS Patients with nosocomial pneumonia from Jan 2003 to Nov 2003 were chosen to be analyzed. RESULTS There were 241 patients with nosocomial pneumonia,the incidence rate of the nosocomial pneumonia in nosocomial infection patients was 41.77%,and the incidence rate of males was much higher than females(1.87∶1),and it was 43.15% in old person among patients with nosocomial pneumonia which was much higher than others,58.72% patients began to have the symptoms in the first 20 days from the day they accessed the hospital,the incidence rate of patients who had been treated by one antibiotic or two antibiotics combination,Immunosuppressors or glucocorticoids drugs were all higher than 50%,the incidence rate of patients who treated by radiotherapy,chemotherapy,thorax-abdomen operations,tracheotomy and ventilator were also quite high,the main pathogens were Gram-negative bacilli(63.95%). CONCLUSIONS We should make heavier prevention and treatment measures in front of target population and their high risk factors and the related rings,so that we can get the real effects.

Lornoxicam is a new non-steroidal anti-inflammatory drug,it can inhibit the activity of cyclooxygenases and the immunologic injury afteroperation.Its efficacies on preoperative,operative period and postoperative analgesia are similar to those of opioid.Lornoxicam can decrease the dosage of opioid drugs and has the good qualities of short half-life,few adverse effects and satisfactory tolerance.Lornoxicam can provide rapid and stable analgesic effect in perioperative phase.

Objective To evaluate the clinical efficacy of transcatheter hepatic arterial thermo-chemotherapy and thermo-lipiodol embolization in the treatment of hepatic metastases from colorectal carcinoma.Methods Sixty-eight cases with hepatic metastases from colorectal carcinoma were equally and randomly divided into two groups.The patients in study group were treated with transcatheter hepatic arterial thermo-chemotherapy and thermo-lipiodol embolization,while the patients in control group were treated with conventional(normal temperature) transcatheter hepatic arterial chemotherapy lipiodol embolization.Results The effective rate of study group and control group was 65%(22 / 34) and 32%(11 / 34) respectively,the difference between two groups was statistically significant(P

Objective:To observe the influence of smoking in the levels of blood lipids and disease activity in the male patients with rheumatoid arthritis(RA), and to provide economical and effective intervention for the RA patients. Methods:A total of 112 male patients with RA were selected.The fasting blood samples from the ulnar vein of all patients were obtained in the morning. The level of C-reaction protein (CRP), erythrocyte sedimentation rate (ESR), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and other indicators were measured. The smoking status (including years of smoking, number of cigarettes smoked per day), tender joint count (TJC), swollen joint count (SJC) and other clinical data were collected and 28-joint disease activity score (DAS28) was calculated.The patients were divided into smoking group (87 cases) and non-smoking group (25 cases).According to the smoking index (SI)(years of smoking multiplied by number of cigarettes smoked per day), the patients were divided into group A (less than 400) (34 cases), group B (between 400 to 800) (37 cases) and group C (more than 800) (16 cases).Results:The levels of TC and TG of the patients in smoking group were significantly higher than those in non-smoking group(P<0.01), the HDL-C level was lower(P<0.01), and there was no significant difference in the LDL-C level between two groups(P>0.05).W ith the increase of SI, the levels of TC and TG were increased(P<0.05), and the HDL-C levels were decreased(P<0.05);the LDL-C levels were increased, but there was no significant difference between various groups(P>0.05).The levels of CRP and ESR in smoking group were higher than those in non-smoking group (P<0.05).The TJC, SJC, and DAS28 of the patients in smoking group were significantly higher than those in non-smoking group(P<0.01).With the increase of SI, the TJC, SJC, and DAS28 were increased(P<0.05), and the CRP and ESR levels showed an increasing trend.Conclusion:Smoking may cause the abnormal blood lipid levels in the patients with RA and the increased disease activity.To quit or reduce smoking may improve blood lipid metabolism in the patients with RA and has a positive effect on reducing the risk of RA.

Objective To investigate the expression of CD4 +CD25 +CD127 low regulatory T cells in peripheral blood mononuclear cells( PBMNCs) in order to explore the influence on the immune status and disease progression in the different period of acute ischemic stroke. Methods 60 patients with acute cerebral infarction were selected as the stroke group,and divided them into the 24~48 h group (n=16), 3~7 d group (n=22) and 8~14 d group (n=22);22 healthy human were set as the control group. To analyze the percentage of CD4 +CD25 +CD127low regu-latory T cells in the peripheral blood of acute cerebral infarction patients and healthy human with flow cytometry. Results The percentages of CD4 +CD25 +CD127 low and CD4 +CD25 high regulatory T cells in the peripheral blood of the stroke group were significantly decreased at 24 ~48 h,3 ~7 d ( P <0.01 ) when compared with the control group, and then tended to return toward the control value at 8~14 d (P>0.05);the percentage of CD4 + regula-tory T cells in the peripheral blood of the stroke group was significantly higher than control group ( P <0.05 ) . Conclusion Imbalance of regulatory T cells is very likely to play an important role in the immunological injury of acute ischemic stroke. Regulatory T cells may be involved in the pathogenesis of acute ischemic stroke,and early detection may provide a basis for treatment.

Objective To explore the value of spectral curve in the differential diagnosis of benign and malignant tumor of the musculoskeletal system.Methods 100 patients with mass of the musculoskeletal system which were found by MRI underwent Dis-covery HD750 Gemstone CT scan ,and the images acquired were processed.Areas with homogenous density which did not contain necrosis,cyst,hemorrhage and calcification were chosen as region of interest(ROI).The optimal Kev on single energy images was selected,then the Kev-CT curve was automatically generated.All 100 patients proved by histological examination were divided into four groups,38 cases were as group A with benign tumor,10 cases as group B with borderline tumor,49 cases as group C with ma-lignant tumor and 3 cases of lipoma(that were analyzed separately since its curve was arc shaped).Average slope rate of every group was calculated according to the formula:K=(HU40keV-HU80keV)/40.Kruskal Wallis test was applied for statistical analysis,P value of less than 0.05 was considered statistical significant.Results The spectral curve of group C was sharply falling type with a mean slope rate of 1.64±1.00,that of group A was gradually falling type with a mean slope rate of 0.75±0.30,and that of group B was 1.34±0.45.The difference of slope rate between group A and C and between A and B were of statistical significance(P<0.05).The spectral curve of 3 lipomas showed arc shaped rising type with a mean slope rate of-2.00.Conclusion Spectral curve is useful in the differential diagnosis of benign and malignant tumor of the musculoskeletal system.Arc shaped curve is a specific sign for tumors containing abundant fat.

BACKGROUND:Excessive mechanical stimulation can lead to endoplasmic reticulum stress. Endoplasmic reticulum stress plays an important role in the occurrence and development of degenerative diseases. Caspase-12 is a specific molecule of endoplasmic reticulum stress, and the intensity of endoplasmic reticulum stress can be reflected by caspase-12 expression. OBJECTIVE:To observe the expression rule of Caspase-12 in chondrocytes under the cyclic tensile stress. METHODS:Human chondrocytes were used as test subjects. After group assignment, mechanical loading system was used. The loading group and corresponding Z-ATAD-FMK inhibitor group received 2, 12, 24, 36, and 48 hours of mechanical stimulation. The blank group (0 hour) and the paired Z-ATAD-FMK inhibitor group received the same process as the loading group except the loading. After loading, cel morphology and growth were observed under the microscope. RT-PCR and western blot assay were used to detect the expression changes of caspase-12 gene and protein in each group. RESULTS AND CONCLUSION:(1) Morphological observation results demonstrated that apoptosis appeared at 2 hours after loading, peaked at 24 hours. With prolonged time, cel growth showed the trend along stress, but apoptosis weakened. It is indicated that cyclic tensile stress could make chondrocyte apoptosis. Endoplasmic reticulum stress might be activated. The model of cel s in vitro was established successful y. (2) Caspase-12 gene and protein expression showed consistent trend, and peaked at 24 hours, which showed significant differences as compared with the blank and inhibitor groups (P<0.05). (3) Cyclic tensile stress can induce chondrocyte endoplasmic reticulum stress, and affect caspase-12 expression.